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HIGHLIGHTS Essential oils from populations of B. dracunculifolia were investigated. β-pinene and (E)-nerolidol were the main compounds in B. dracunculifolia populations. The difference in the chemical profile of the essential oils is quantitative only. There is a negative correlation between the antioxidant activity and spathulenol.

Haemonchus contortus, a blood-sucking parasite of small ruminants, produces very important economic losses in the productive sector. This abomasum parasite has become resistant to most commercial drugs worldwide, and alternatives to fight this problem are urgently needed. Essential oils (EO) are a complex mixture of volatile secondary metabolites, composed mainly by terpenoids and phenolic compounds, from plants that have several pharmacological properties, including anthelmintic activity. Particularly, citrus peel is a source of cold-pressed EO, where limonene is its major component, and can be used as an additional food component for ruminants. The aim of the present work was to determine the in vitro anthelmintic activity of EO from Citrus bergamia (EOB), C. x paradisii (EOG) and limonene against the benzimidazole-susceptible Kirby isolate of H. contortus, using the egg hatch test (EHT) and the exsheathed third stage larval motility test (XLMT) using a WMicroTracker equipment. Albendazole (ABZ) and monepantel (MON) were used as positive controls. The 50% inhibitory concentrations (IC50) in XLMT were 8.77 and 13.88 µg/ml for EOB and EOG respectively, after an incubation of 72 h. An interesting observation on XLMT resulted when the positive controls were tested on the same plate, but in different well of the EOB. The volatile components of the EO significantly influenced (P

The essential oil produced by Eugenia uniflora leaves is intensively used by the cosmetic, pharmaceutic and perfumery industries mainly because its antioxidant effect and antimicrobial and antihelmintic ativities. Drying and storage of the aromatic plant leaves can affect the yield and quality of its essential oil. This study aimed to evaluate the effect of drying method and storage conditions on the essential oil yield and composition in Eugenia uniflora leaves. The essential oil were extracted from dried leaves at ambient temperature and at 45°C for six drying periods (0, 6, 24, 48, 72 and 96 hours-h). The essential oil yield was also analysed after 0, 60, 120, and 180 days of storage in three types of packaging. The oil was extracted by hydrodistillation and the composition was determined by gas chromatography-mass spectrometry. Results showed that drying the leaves at ambient temperature and 45°C for 48 and 24 h, respectively, increased the essential oil yield. At 45°C the monoterpene and sesquiterpene compounds increased, with curzerene+viridiflorene identified as the major compounds. Package types did not affect the essential oil yield but the increase of the storage period decreased it.

Lucilia cuprina, known as the Australian blowfly, is of high medico-sanitary and veterinary importance due to its ability to induce myiasis. Synthetic products are the most frequent form of fly control, but their indiscriminate use has selected for resistant populations and accounted for high levels of residues in animal products. This study aimed to assess the effect of essential oil from leaves of Curcuma longa (CLLEO), and its major compound α-phellandrene against L. cuprina L3. An additional goal was to determine the morphological alterations in target organs/tissues through ultrastructural assessment (SEM) and light microscopy, as well as macroscopic damage to cuticle induced by CLLEO. Groups of 20 L3 were placed on filter paper impregnated with increasing concentrations of CLLEO (0.15 to 2.86 μL/cm2) and α-phellandrene (0.29 to 1.47 μL/cm2). Efficacy was determined by quantifying L3 mortality 6, 24 and 48 h after contact with CLLEO and by measuring the structural damage to L3. CLLEO and α-phellandrene inhibited adult emergence by 96.22 and 100%, respectively. Macroscopic cuticle damage, appeared as diffuse pigment and darkening of larval body, was caused by both extracts. The SEM revealed dryness on the cuticle surface, distortion of the sensorial structures and general degeneration in treated L3. Furthermore, alterations in target organs (digestive tract, fat body and brain) were noticed and shall be used as biomarkers in future attempts to elucidate the mechanism of action of these compounds. The vacuolar degeneration and pyknotic profiles observed in the brain tissue of treated larvae with both extracts and the decreased motility within

The restinga is an Atlantic Forest ecosystem characterized by tree, shrub, and herb species that are rich sources of essential oils. In this study, we aim to quantify the essential oil content and determine the chemical constituents of fresh leaves of 14 plant species in a restinga stretch in southern Brazil. Essential oils were obtained by hydrodistillation in a Clevenger-type apparatus and analyzed by gas chromatography coupled to mass spectrometry. Campomanesia reitziana, Cortaderia selloana, and Sophora tomentosa had no essential oils. Total essential oil content ranged from 0.01% (Mikania involucrata) to 1.56% (Varronia curassavica). In total, 60 chemical constituents were identified, representing between 46.2% and 96.5% of the chemical composition of the essential oils. Limonene was the common constituent in all species in which the essential oils were present. The major constituents were ar-curcumene (15.1%) and cis-chrysanthenol (14.2%) in Ambrosia elatior; benzyl benzoate (43.5%) and benzyl salicylate (23.7%) in Aniba firmula; caryophyllene oxide (35.7%) and spathulenol (10.6%) in Austroeupatorium inulaefolium; spathulenol (19.8%) and caryophyllene oxide (14.0%) in Baccharis spicata; caryophyllene oxide (16.3%) in Eugenia astringens; curzerene (30.0%), limonene (13.0%), and germacrone (11.9%) in Eugenia uniflora; caryophyllene oxide (17.1%) and ledol (11.3%) in Lantana camara; caryophyllene oxide (27.7%) and limonene (12.7%) in M. involucrata; 1,8-cineole (19.8%) in Psidium cattleianum; limonene (10.2%) in Schinus terebinthifolius, and allo-aromadendrene (15.2%) in V. curassavica. We expect that our results can assist in selecting species of potential interest for herbal, phytotherapeutic, and cosmetic products.

Varronia curassavica Jacq. (Boraginaceae) is a native species of the Atlantic Forest with medical importance. This study determined the essential oil variation of 16 populations of V. curassavica of restinga in Santa Catarina, Brazil, in response to drying and edaphoclimatic conditions. The populations supplied essential oil with a content between 0.27 to 1.15% in the summer and from 0.33 to 1.12% in the winter. 41 chemical compounds were identified in the summer and 40 in the winter. The compounds were grouped into 4 chemical classes in each station studied. The common chemical constituents found in the essential oil of all populations and in both seasons were α-thujene, α-pinene, sabinene, α-humulene, (E)-cariophylene, spatulenol, mircene, allo-aromadendrene, β-sesquifelandreno and α-zingiberene. Cluster analysis using the nearest neighbor method based on Euclidean distance grouped the 16 populations into 3 groups in the summer and 8 groups in the winter. As the habitats have distinct pedological characteristics, we identified that pH, organic matter, sum of bases and base saturation are associated with the synthesis of (E)-caryophyllene, α-humulene, and allo-aromadendrene from populations.

The weevils Sitophilus oryzae L. and Sitophilus zeamais Motsch. are stored-product pests that are mainly controlled with synthetic insecticides. Insecticides made from essential oils (EOs) from bioactive plants can provide alternative control options that are safer and less environmentally damaging. This study investigated the chemical composition and insecticide activity of the EOs of Tagetes minuta L., Baccharis uncinella DC, Baccharis dracunculifolia DC, Salvia officinalis L., Mentha arvensis L., and Melaleuca alternifolia C. against S. oryzae and S. zeamais. The number of compounds we identified in the EOs studied ranged between 10 for T. minuta and 21 for B. uncinella. Only the EOs of T. minuta and M. arvensis showed insecticidal activity. Fumigation was more effective than the contact method for both EOs. The EO of M. arvensis had the highest level of toxicity in the contact method, with an LC50 of 71.58 μL/L for S. oryzae and 67.31 μL/L for S. zeamais. It also had the highest toxicity in the fumigation method (LC50 of 56.41 μL/L for S. oryzae and 47.03 μL/L for S. zeamais) with the lowest lethal time (LT50 of 8.9 h for S. oryzae and 8.1 h for S. zeamais). The EO of M. arvensis, therefore, has the greatest potential for development as a new botanical insecticide for the control of stored-product adult weevils.

The present research deals with the collection and structural elucidation of an unknown component, accounting for about 35% of the essential oil obtained upon distillation of the leaves of Eugenia uniflora L., harvested during summer (January, 2017) in Paraná State (Southern Brazil). A multidimensional gas chromatographic preparative system, based on the coupling of three GC systems equipped with apolar, PEG and ionic liquid-based stationary phases, was successfully applied for the isolation of the chromatographic band relative to the unknown molecule. The use of wide-bore columns allowed for an increased sample capacity compared to conventional micro-bore columns, thus the injection of a neat sample was feasible, greatly reducing the total collection time. A higher chromatographic efficiency was afforded by the use of a multidimensional approach in the heart-cut mode, exploiting the different selectivity of three stationary phases, which ensured the attainment of a highly pure fraction. In only five runs, more than 3 milligrams were collected, with an average purity greater then 95%. Finally, the unknown component was subjected to nuclear magnetic resonance spectroscopy, mass spectrometry and condensed phase Fourier-transform infrared spectroscopy, leading to the identification of 6-ethenyl-6-methyl-3,5-di(prop-1-en-2-yl)cyclohex-2-en-1-one. The presented approach has been demonstrated to be effective for the isolation and structural elucidation of unknown molecules in complex samples, which will allow for further in-depth studies, like biological evaluation or pharmacological tests.

Eugenia uniflora, known as the “Brazilian cherry”, is an economically important neotropical Myrtaceae in the cosmetics and pharmaceutical industries due the production of essential oils with antioxidant activity. On account of its significant genetic variability, genotype evaluations are needed in order to identify genetic features related to the essential oil production that meet the industry requirements. The main objective of the present study was to evaluate the yield, composition, and antioxidant activity of essential oils isolated from the leaves of 36 genotypes of E. uniflora. Essential oil samples were obtained by hydrodistillation, and their composition was determined by gas chromatography coupled with mass spectrometry. A variation of 0.22% to 1.68% in the essential oil yield was observed, in which 78 compounds, namely oxygenated sesquiterpenes, were identified. According to the cluster analysis of the major compounds, six groups were revealed. The observed diversity demonstrates the genetic variability of the species. Also, the antioxidant activity was affected by the composition of the essential oils, ranging from 176.66 to 867.57 µM TEAC.

The present work was conducted to evaluate the effects of Tributilin (TBT) on chromosomes of the Neotropical fish Astyanax sp. Twenty nine individual fishes (8 females and 21males) were divided into two groups. Within 15 fishes in one group, 7 fishes were treated with TBT for 19 d, while other 8 fishes were without, and were sacrificed for the observation. Another 14 fishes (8 for treatment, and 6 as control) were treated for 37d. Chromosomal aberration was detected in both group of treated and of non-treated fishes. However, the frequency of aberration of chromosome was significantry higher in the treated fishes. These data represent the first evidence of the effects of TBT on chromosomes of a Neotropical fish species.

Cytogenetic studies have assisted in the taxonomic classification of organisms, especially those involving species with highly similar morphologic characteristics, or so-called cryptic species. Strongylura marina and Strongylura timucu collected from Paranagua Bay, Parana Coast in Southern Brazil are considered cryptic species, and the identification of interspecific variations based on the number and/or morphology of its chromosomes may serve as differentiating cytotaxonomic markers. Chromosomes of the two species were subjected to different banding and staining methods (C-, Ag-, and DAPI-CMA3), as well as chromosomal mapping of major rDNA (45S), revealed with an 18S probe by fluorescence in situ hybridization (FISH). The pattern of distribution of constitutive heterochromatin showed distinct features involving the pericentromeric and telomeric bands in both species. In S. marina, chromosome 1 represents the main species-specific marker, appearing almost entirely heterochromatic. In both species, the 45S rDNA is located at terminal region of the short arm of the chromosome 6, as detected by silver nitrate staining and FISH. Despite the apparent conserved diploid number of 48 chromosomes, data on the karyotype microstructure characterize the cytogenetic profile of the genus and may allow the establishment of cytotaxonomic and evolutionary inferences for these fishes.

In this study, five species of marine fishes from the Paranaguá Bay in the Brazilian coast were evaluated. Eucinostomus argenteus and Diapterus rhombeus (Gerreidae) presented 48 chromosomes, all of which more acrocentric (FN = 48); Strongylura timucu and S. marina (Belonidae) also presented 48 chromosomes, but with a higher karyotypic complexity than the Gerreidae, 10M+2SM+36A (FN = 60) and 4M+44A (FN = 52), respectively. The fifth species, Mugil curema (Mugilidae), different than the others, presented only 28 chromosomes 20M+4ST+4A (FN = 48). The species presented diversity in the karyotypic macro-structure, which should be relevant for the cytotaxonomy and the evolution of this group of the vertebrate.Nas últimas décadas tem ocorrido no Brasil um incremento de estudos cariotípicos em peixes marinhos. Atualmente são conhecidos os cariótipos de 118 espécies, distribuídas em 43 famílias e 80 gêneros. Foram estudadas cinco espécies de peixes marinhos do complexo estuarino da Baía de Paranaguá na costa brasileira. Eucinostomus argenteus e Diapterus rhombeus (Gerreidae), apresentaram 48 cromossomos todos acrocêntricos (NF = 48); Strongylura timucu e S. marina (Belonidae) apresentaram 48 cromossomos, porém com complexidade cariotípica maior do que apresentada pelos gerreídeos, 10M+2SM+36A (NF = 60) e 4M+44A (NF = 52), respectivamente. A quinta espécie, Mugil curema (Mugilidae), ao contrário das outras quatro espécies aqui analisadas, apresentou apenas 28 cromossomos 20M+4ST+4A (NF = 48). Apesar da tendência em se verificar um cariótipo constituído por 48 cromossomos em teleósteos marinhos, as espécies aqui analisadas apresentam uma diversidade para a macroestrutura cariotípica a ser considerada para a citotaxonomia e evolução desse grupo de vertebrados.

Chromosomal analyses were performed in the fish Astyanax sp.D collected from three different points: two streams from the right bank and one from the left bank of the Upper Iguaçu River, Paraná State, Brazil. The individuals from all localities possess 2n = 50 chromosomes and a FN = 84 (4m+24sm+6st+16a). The C-banding pattern was similar in all populations. However, within each population, an interindividual variation concerning the number and localization of heterochromatic bands was observed. Some of these variations were quantified in each population, and the results indicate that the samples were not different when studying the variable frequencies. Considering that Astyanax sp.D is typical in the headwaters of the Iguaçu River, these results were not expected. The data indicate that gene flow is occurring and that the Iguaçu River is not an ecological barrier among the Astyanax sp. D populations.

Chromosomal features, location and variation of the major and minor rDNA genes cluster were studied in three pufferfish species: Sphoeroides greeleyi and Sphoeroides testudineus (Tetraodontidae) and Cyclichthys spinosus (Diodontidae). The location of the major rDNA was revealed with an 18S probe in two loci for all species. The minor rDNA loci (5S rDNA) was found in one chromosome pair in tetraodontid fishes and four sites located on two distinct chromosomal pairs in C. spinosus. A syntenical organization was not observed among the ribosomal genes. Signal homogeneity for GC/AT-DNA specific fluorochromes was observed in diodontid fish except in the NORs regions, which were CMA3-positive. Giemsa karyotypes of tetraodontid species presents 2n = 46, having the same diploid value of other Sphoeroides species that have been investigated. On the other hand, the karyotype of C. spinosus, described for the first time, shows 2n = 50 chromosomes (4m + 18sm + 12st + 16a). The foreknowledge of the karyotypic structure of this group and also the physical mapping of certain genes could be very helpful for further DNA sequence analysis.

Karyotype of M. curema from the Gulf of Mexico and Brazil have been reported as possessing chromosome complement with 2n=28 and FN=48, whereas specimens from Venezuela has been reported as possessing a diploid number 2n=24 and a conserved FN (48). Although at first sight this variation suggests the presence of a chromosomal intraspecific (interpopulational) variability, the possibility that we are dealing with two different species was examined. This work revisit the karyotypes of M. curema from Venezuela and Brazil, including new data on C-banding, and NOR localization, and compares morphologic characteristics of samples from both localities. Thus, besides diploid number, the constitutive heterochromatin distribution and NORs location, mark other differences between M. curema Cytotype 1 (2n=28; FN=48) and Cytotype 2 (2n=24; NF=48). Moreover, morphologic comparison revealed differences in the scale counts and pectoral fin rays: 35 scales in the middle body line and 15 pectoral fin rays in specimens possessing the karyotype 2n=28, compared with 37-39 scales in the middle body line and 17 pectoral fin rays in specimens with the karyotype 2n=24. These differences lead us to suggest that both cytotypes are not related merely to geographic polytipic variations but could correspond to different species. Os cariótipos de M. curema do Golfo do México e do Brasil possuem 2n=28 cromossomos e NF=48. Espécimes da Venezuela, entretanto, apresentam um número diplóide de 28 cromossomos e um NF conservado (48). Apesar desta variação sugerir, a princípio, uma variabilidade intraespecífica (interpopulacional), a possibilidade de estarmos perante diferentes espécies foi investigada. O presente trabalho re-analisa os cariótipos de M. curema da Venezuela e do Brasil, incluindo novos dados sobre bandamento C e localização das NORs e compara caracteres merísticos e morfométricos de amostras de ambas as localidades. Assim, junto com o número cromossômico, a heterocromatina constitutiva e a distribuição das NORs trazem outras diferenças entre M. curema Citótipo 1 (2n=48, FN=48) e Citótipo 2 (2n=48, FN=48). Além disso, as comparações dos caracteres morfológicos revelam diferenças nas contagens, com 35 escamas na linha média do corpo e 15 raios nas nadadeiras peitorais nos espécimes com cariótipo 2n=28, já o cariótipo 2n=24 apresenta 37-39 escamas e 17 raios nas peitorais. Essas diferenças sugerem que ambos citótipos não estão relacionados meramente a variações geográficas politípicas mas que podem corresponder a diferentes espécies.